Smartphone and handheld fluorometer enable rapid point of care testing of Escherichia coli in urinary tract infections via specific proteolytic cleavage and cascade amplifications

Culture-free, rapid, and ultrasensitive quantification of Escherichia coli (E. coli) is urgently needed to diagnose clinical urinary tract infections (UTIs), especially point-of-care testing (POCT). Herein, a homogeneous sensitive multi-modal (fluorometer, smartphone, and self-developed handheld fluorometer) assay was developed to detect E. coli within 30 min, and nanoscale luminescent cadmium telluride quantum dots (CdTe QDs) were employed as signal reporters. E. coli outer membrane protease T (OmpT) is a biomarker for the specific proteolytic cleavage of peptides. p-Aminophenol (PAP), generated by leucine aminopeptidase (LAP) via secondary cleavage, was used to reduce Ag+. Cascade amplifications included Ag+ and copper sulfide nanoparticles (CuS NPs) to release Cu2+, CdTe QDs, and Cu2+/CuS NPs, which can ensure detection sensitivity. The method achieved single bacterium detection with great stability and selectivity. The fluorometer, handheld fluorometer and smartphone red-green-blue (RGB) value-based modes were determined to be 50, 15, and 15 clinical urine samples, respectively. The findings were consistent with clinical culture counting, and an area under the curve (AUC) of 0.95 with 90% sensitivity and 100% specificity was achieved. Overall, this OmpT-based catalytic biosensor can provide a solution for the POCT diagnosis of UTIs based on E. coli detection. •A homogeneous rapid fluorescence one-pot assay for Escherichia coli was developed.•The non-culture, rapid, multi-modal sensor had high sensitivity and selectivity.•This system was successfully applied to the analysis of E. coli in 50 urine clinical samples.